Faculty of Medicine
Library of the School of Health Sciences

2021-12-01

2021

Zevolis Evangelos

Κουτσιλιέρης Μιχαήλ, Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ
Τρουπής Θεόδωρος, Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ
Κασσή Ευανθία, Καθηγήτρια, Ιατρική Σχολή, ΕΚΠΑ
Φιλίππου Αναστάσιος, Αναπληρωτής Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ
Μαυραγάνη Κλειώ, Αναπληρώτρια Καθηγήτρια, Ιατρική Σχολή, ΕΚΠΑ
Αγγελογιάννη Παναγούλα, Αναπληρώτρια Καθηγήτρια, Ιατρική Σχολή, ΕΚΠΑ
Χατζηγεωργίου Αντώνιος, Επίκουρος Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ

Ο ρόλος της μηχανικής φόρτισης στην υπερτροφία και στη φλεγμονή του μυοκαρδίου: In vitro μελέτες

Greek

The role of mechanical stretch in myocardial hypertrophy and inflammation: in vitro studies

Background: Cardiomyocytes possess the ability to respond to mechanical stimuli by adapting their biological functions. H9C2 cardiomyoblasts is a cell line used as an alternative for cardiomyocytes. They are used in vitro as a mimetic model for cardiac muscle due to their biochemical, morphological, and hormonal signalling properties. Thus, several studies have used H9C2 cells for examining their differentiation features towards a cardiac-like phenotype. This study investigated the adaptations of H9C2 cells during differentiation and the effects of different loading protocols on signalling and gene expression responses of myogenic, anabolic, inflammatory, atrophy and pro-apoptotic factors in differentiated cardiomyocyte-like H9C2 cells. Methods: H9C2 cardiomyoblasts cultured and differentiated in order to investigate differentiation process. Undifferentiated Η9C2 cardiomyoblasts used as control. Afterward, differentiated H9C2 cells were cultured on silicon elastic membranes and then underwent passive, cyclic mechanical stretch. Flow cytometry was used to characterize cell cycle of the H9C2 cardiomyoblasts and immunofluorescence to monitor structural changes during their differentiation. Moreover, alterations in the mRNA and protein expression levels of cardiac specific markers, as well as of cell cycle regulatory factors, myogenic regulatory factors (MRFs), IGF-1 isoforms, apoptotic, atrophy and inflammatory factors were evaluated during differentiation. p-Erk 1/2 and p-Akt, the myogenic factors MyoD and Myogenin and apoptotic factor p53 were determined by immunoblotting from stretched and non-stretched cardiomyocytes. qRT-PCR was used to measure changes in expression levels of the myogenic regulatory factors, growth, apoptotic, atrophy and inflammatory factors in response to mechanical loading of the H9C2 cells. Results: Compared to undifferentiated cells, differentiated cardiomyocyte-like H9C2 cells (5th day of differentiation) exhibited increased expression of cardiac specific markers, myosin heavy chain, MRFs, and IGF-1 isoforms. Moreover, cells that underwent a 12 hours duration mechanical stretch protocol increased expression of IGF-1 isoforms and decreased expression of apoptotic, atrophy and inflammatory factors, compared both to mechanical loading protocols with duration 24 hours and 15 minutes and to unloaded cells. Additionally, low frequency (0.25 Hz)/strain (2%) mechanical stretching protocol showed an enhanced expression of MRFs and IGF-1 isoforms along with a decreased expression of apoptotic, atrophy and inflammatory factors, compared both to high strain/frequency mechanical loading protocols and to unloaded cells. Conclusions: The findings of this study suggest that cardiomyoblast H9C2 exhibit a myogenic and cardiac differentiation at their 5th day of differentiation. Also, the effects of mechanical stretching on these differentiated cardiomyocyte-like cells suggest that a 12 hours duration, low frequency (0.25 Hz)/strain (2%) stretching protocol can be overall the most effective in inducing beneficial responses in these cardiomyotubes.

Health Sciences

Cardiomyocytes, Cellular mechanotransduction, H9C2, Mechanical stretch, Myocardiac hypertophy

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https://pergamos.lib.uoa.gr/uoa/dl/object/2967623